In acute promyelocytic leukaemia (APL), the promyelocytic leukaemia (PML) protein is fused to the retinoic acid receptor α (RAR). This disease can be treated effectively with arsenic, which induces PML modification by small ubiquitin-like modifiers (SUMO) and proteasomal degradation. Here we demonstrate that the RING-domain-containing ubiquitin E3 ligase, RNF4 (also known as SNURF), targets poly-SUMO-modified proteins for degradation mediated by ubiquitin. RNF4 depletion or proteasome inhibition led to accumulation of mixed, polyubiquitinated, poly-SUMO chains. PML protein accumulated in RNF4-depleted cells and was ubiquitinated by RNF4 in a SUMO-dependent fashion in vitro. In the absence of RNF4, arsenic failed to induce degradation of PML and SUMO-modified PML accumulated in the nucleus. These results demonstrate that poly-SUMO chains can act as discrete signals from mono-SUMOylation, in this case targeting a poly-SUMOylated substrate for ubiquitin-mediated proteolysis.
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This work was supported by Cancer Research UK and the RUBICON EU Network of Excellence. A. P was supported by a Wellcome Trust Studentship. We would like to thank Hugues de The for helpful discussions and providing the chicken anti-PML antibody, and Douglas Lamont, manager of the Fingerprints Proteomics Facility, University of Dundee, for generating the Orbitrap mass spectrometry data. The provision of critical reagents by Roel van Driel and Dan Bailey is gratefully acknowledged.
The authors declare no competing financial interests.
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Tatham, M., Geoffroy, MC., Shen, L. et al. RNF4 is a poly-SUMO-specific E3 ubiquitin ligase required for arsenic-induced PML degradation. Nat Cell Biol 10, 538–546 (2008). https://doi.org/10.1038/ncb1716
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